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目的研究survivin反义寡核苷酸对肾透明细胞癌786O细胞表达survivin蛋白、细胞凋亡、增殖的影响。方法设计并合成特异性靶向survivin的反义寡核苷酸(ASODN)。肾透明细胞癌细胞株786O分为6组:空白对照组、脂质体转染对照组、正义链转染对照组、200、400和600nmol/LASODN转染组。作用24h后收获各组细胞。倒置显微镜及透射电子显微镜观察细胞形态变化,免疫组化法检测各组细胞survivin表达情况和流式细胞术检测各组细胞周期变化和凋亡指数,MTT法检测survivin反义寡核苷酸对各组细胞的生长抑制率。结果电镜下可以见到典型凋亡样改变,而各对照组细胞生长良好;各ASODN转染组细胞survivin表达有不同程度减弱;各ASODN转染组细胞凋亡指数明显高于各对照组(P<0.05),以600nmol/LASODN转染组最为明显(P<0.05),而各对照组间差异无显著性(P>0.05)。结论不同浓度survivinASODN转染后能下调survivin蛋白表达,诱导肾透明细胞癌786O细胞凋亡,抑制786O细胞增殖。
Objective To investigate the effect of survivin antisense oligonucleotide on the expression of survivin protein, apoptosis and proliferation in renal clear cell carcinoma 786O cells. Methods Antisense oligonucleotides (ASODNs) targeting specifically to survivin were designed and synthesized. The renal clear cell carcinoma cell line 786O was divided into 6 groups: blank control group, lipofection control group, sense strand transfected control group, 200,400 and 600 nmol / L ASODN transfection group. The cells in each group were harvested after 24 hours. The morphological changes of cells were observed by inverted microscope and transmission electron microscope. The expression of survivin in each group was detected by immunohistochemistry and the cell cycle and apoptosis index were detected by flow cytometry. The expressions of survivin antisense oligonucleotides Group cell growth inhibition rate. Results Apoptosis-like changes were observed under electron microscope, and the cells in each control group grew well. The expression of survivin in ASODN-transfected cells was attenuated to varying degrees. The apoptosis index in each ASODN transfected group was significantly higher than that in control group (P <0.05). The most significant difference was found in 600 nmol / L ASODN transfection group (P <0.05), but there was no significant difference among the control groups (P> 0.05). Conclusions Survivin ASODN can down - regulate the expression of survivin protein and induce the apoptosis of 786O cells and inhibit the proliferation of 786O cells.